Setareh Zahedian - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Azadeh Hekmat - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Saeed Hesami Tackallou - Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran.
Mahmood Ghoranneviss - Department of Plasma Physics Research Center, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Background: For many years, the chemotherapeutic agent doxorubicin (DOX) has been used to treat various cancers; however, DOX initiates several critical adverse effects. Many studies have reported that non-thermal atmospheric pressure plasma can provide novel, but challenging, treatment strategies for cancer patients. To date, tissues and cells have been treated with plasma-activated medium (PAM) as a practical therapy. Consequently, due to the harmful adverse effects of DOX, we were motivated to elucidate the impact of PAM in the presence of DOX on MCF-۷ cell proliferation. Methods: MTT assay, N-acetyl-L-cysteine (NAC) assay, and flow cytometry analysis were utilized in this research.  Results: The results demonstrated that ۰.۴۵ μM DOX combined with ۳-min PAM significantly induced apoptosis (p< ۰.۰۱) through intracellular ROS generation in MCF-۷ when compared with ۰.۴۵ μM DOX alone or ۳-min PAM alone. In contrast, after treatment with ۰.۴۵ μM DOX plus ۴-min PAM, cell necrosis was increased. Hence, DOX combined with ۴-min PAM has cytotoxic effects with different mechanisms than ۴-min PAM alone, in which the number of apoptotic cells increases. Conclusions: Although further investigations are crucial, low doses of DOX plus ۳-min PAM could be a promising strategy for cancer therapy. The findings from this research may offer advantageous and innovative clinical strategies for cancer therapy using PAM.

Apoptosis, Breast cancer lymphedema, Doxorubicin, Plasma-activated medium (PAM), Necrosis.